92 results about "PHA binding" patented technology

The invention relates to the technical field of biological detection, and discloses a method for double-ring detection of lung cancer L858R mutation based on CRISPR / Cas9 and DNA, and the method comprises the following steps: enriching circulating tumor cells CTC from a blood sample; extracting genomic DNA (deoxyribonucleic acid) of the enriched CTC; carrying out amplification on the DNA fragment containing the L858R mutation by adopting recombinase polymerase amplification RPA (recombinase polymerase amplification); the amplified DNA fragment is hybridized with a recognition ring in a DNA double-ring structure, a DNA double ring is composed of the recognition ring and a report ring, and a part of region of the recognition ring is hybridized with a corresponding fragment of the report ring to form a staggered double-chain region; when L858R mutation exists, the mutated DNA and a recognition ring are hybridized to activate a CRISPR / Cas9 system, and under the guidance of sgRNA, Cas9 cuts hybrid double strands and releases a report ring; carrying out rolling circle amplification RCA under the action of DNA polymerase by taking the released report ring as a template; a fluorescent probe is used for detecting an RCA amplification product, and whether L858R mutation exists or not is judged according to a fluorescence signal. The CRISPR / Cas9 system is combined with a DNA double-ring structure, so that the ultra-sensitive and specific detection on the lung cancer L858R mutation is realized.

The present application relates to an anti-CD3 antibody and its application. The present application develops a specific antibody molecule targeting CD3 epsilon chain, which can activate T cells by combining with CD3 epsilon on T cells. A T cell binding protein containing the CD3 antibody is further designed, which can specifically target and bind to CD3 epsilon on the surface of T cells in vivo and in vitro, so as to achieve moderate activation of T cells. The CAR lentivirus containing the T cell binding protein of the present application can induce the generation of CAR-T cells in vitro and in vivo, and the CAR-T cells have good tumor cell killing efficiency.

The invention relates to a method for splitting, assembling and site-specific integration of large-fragment exogenous DNA (deoxyribonucleic acid) in Trichoderma reesei. Comprising the following steps: (1) constructing an arg:: Ptcu-Cas9-TtrpC-hph strain containing a Cas9 protein; (2) constructing a pUC19 <-> integration site: a pyr4 plasmid; (3) obtaining a plurality of target gene segments containing integration site homologous arms and having overlapping regions at the tail ends of adjacent DNA segments; (4) obtaining a gRNA expression cassette of a targeted genome integration site; (5) arg conversion: the arg is converted into a Ptcu-Cas9-TtrpC-hph strain; and (6) screening and verifying. According to the method, the CRISPR / Cas9 system is combined with the TAR in the trichoderma reesei for the first time, the exogenous large-fragment DNA is split into a plurality of small fragments, then the plurality of small-fragment exogenous DNA are assembled in the trichoderma reesei and are subjected to fixed-point integration, one-step editing of the large-fragment exogenous DNA is realized, and compared with a traditional genetics method, the gene editing efficiency in the trichoderma reesei is greatly improved.

It is provided an anti-Aβ34 antibody and its use for diagnosing Alzheimer's disease in a patient, comprising obtaining a sample from the patient, detecting the level of Aβ34 in the sample by contacting the sample with the anti-Aβ34 antibody and detecting binding between Aβ34 and the antibody, and diagnosing the patient with Alzheimer's disease when the presence of Aβ34 in the sample is detected, alone or in combination with detecting an amyloid deposition marker such as Aβ42 and determining the ratio of Aβ34 / Aβ42.

The present application relates to the technical field of RNA extraction, and particularly relates to a pretreatment reagent, a method and a kit for RNA purification of whole blood samples. By introducing hydrophobic DESs as the core pretreatment component, the DESs are mixed with the whole blood sample in advance, the DESs form an independent phase layer after centrifugation by using the unique physicochemical properties, and the liposoluble drug components, inflammatory proteins and pigments are selectively extracted to the hydrophobic DESs phase and the RNA is preliminarily purified; the upper water phase of the pretreatment and purification is taken and added into a lysis solution containing a chaotropic salt to fully release the nucleic acid, and DNase I is introduced for digestion. The present application combines the pretreatment of hydrophobic DESs and the digestion of DNase I, and is used in combination with PVP (targeting adsorption of residual pigments and phenolic compounds), reduces the impurities of the lysis system through the front-end pretreatment, creates a low-interference and high-efficiency purification environment for the specific binding of RNA by the magnetic beads, and forms a synergistic removal network for the interference of complex samples.

The application belongs to the technical field of biotechnology, and discloses a nano antibody targeting B cell BCMA and application thereof. The amino acid sequences of the complementarity determining regions CDR1-CDR3 of the nano antibody are selected from SEQ ID NO. 1-12. The nano antibody provided by the application can specifically bind to B cells in human blood, especially activated plasma cells, so as to eliminate B cells producing pathological antibodies. Compared with traditional antibodies, the nano antibody provided by the application has a smaller molecular weight, high stability, low cost, easy expression and high expression amount, and is suitable for large-scale production. The nano antibody is combined with a solid-phase carrier in the application, so that B cells in blood can be targeted and eliminated, the number of B cells in blood of patients can be reduced in a short time, and the antibody level can be reduced, thereby improving the quality of life, reducing complications, and providing a new treatment option for patients who are ineffective or intolerant to traditional treatment.

The present application relates to a kind of sgRNA targeting Plagl1 gene DMR, epigenetic editing system and its application.Specifically, the present application provides sgRNA targeting Plagl1 gene differential methylation region, which forms epigenetic editing system with dCas9-Tet1-CD fusion protein.The system is loaded in injectable type light-cured silk fibroin hydrogel, the expression of Plagl1 gene in periosteum stem / progenitor cells is accurately activated by targeted demethylation modification, thereby promoting osteogenic differentiation.The present application combines epigenetic regulation with biomaterial delivery, and provides a safe, efficient new strategy for jaw bone defect regeneration and repair.

The invention discloses a solid-phase and liquid-phase combined synthesis method of tilpotide, and belongs to the field of chemical synthesis of polypeptide drugs. The method comprises the following steps: firstly, synthesizing a complete main peptide chain of the tiall peptide on a solid phase carrier, and protecting a side chain of lysine (Lys24) at the 24th site by adopting an orthogonal protecting group (Dde), so that side chain activated ester (Ecosanoic acid-gamma-Glu-AEA-AEA-OSu) can be specifically connected to an amino group of lysine (Lys20) at the 20th site, and the protecting group of the Lys24 can be selectively removed under a mild liquid phase condition to obtain crude peptide of the tiall peptide. Complex side chain modification is transferred from a solid phase to a liquid phase, the problems of steric hindrance and low connection efficiency are effectively solved, and the method has the advantages of being high in synthesis purity, few in side reaction, easy to amplify and the like and is suitable for industrial production.

The invention belongs to the technical field of biological medicines, and relates to a mutants of mu-conotoxin KIIIA rich in positive charges and application of the mutants. The mutant is obtained by implementing site-specific mutagenesis at the following positions on a mutant as shown in SEQ ID NO.1 in a sequence table: S6R; the amino acid sequence of the mutant is as shown in SEQ ID NO. 2 in a sequence table. According to the invention, systematic structural optimization is carried out on a parent peptide KIIIA containing three pairs of disulfide bonds, and a strategy of combining disulfide bond deletion and amino acid site-directed mutagenesis is adopted, so that a mutant which is simple and convenient to synthesize and has a remarkable inhibition effect on a NaV1.4 channel is obtained. The mutant has high activity, high stability and better pharmacokinetic characteristics, and has wide development potential.

Exemplary systems, methods, and computer-accessible medium are provided for Chimeric Antigen Receptor (CAR)-T therapy. Thus, the exemplary systems, methods, and computer-accessible medium are provided that select a first set of targets highly expressed in cancer and lowly in normal tissues, select a second set of targets that are highly co-expressed in the cancer and lowly in normal tissues, generate an input set by combining the first and second set of targets with dependency data, single-cell heterogeneity, and antibody availability, determine, by a learning model, at least one target pair for use in CAR-T therapy, and apply a cell therapy based on the determined target pair(s).

This invention belongs to the field of biomedical technology, specifically relating to a peptide targeting TRPV1 and its applications. A peptide targeting TRPV1 has the amino acid sequence shown in SEQ ID NO.1. The peptide also includes a mutant obtained by amino acid scanning mutation based on the sequence shown in SEQ ID NO.1. The amino acid sequence of the mutant is shown in any one of SEQ ID NO.2-16. This peptide is used to prepare formulations that block or inhibit TRPV1 activity. This invention provides a peptide targeting TRPV1 and its applications. Through a strategy combining artificial intelligence screening and structural prediction, SEQ ID NO.1 was successfully identified and its mechanism of action was elucidated. Experimental results show that SEQ ID NO.1 and its mutants exhibit excellent analgesic activity and good safety both in vitro and in vivo, demonstrating significant clinical application potential.

The invention belongs to the technical field of biological medicines, and relates to a mutants of mu-conotoxin KIIIA rich in positive charges and application of the mutants. The mutant is obtained by carrying out site-directed mutagenesis on C1A, K7R and C15R on a natural mu-conotoxin KIIIA sequence; the amino acid sequence of the mutant is shown as SEQ ID NO. 1 in a sequence table. According to the invention, a series of mutants which are simple and convenient to synthesize and have a remarkable inhibition effect on a NaV1.4 channel are obtained by performing systematic structural optimization on a parent peptide KIIIA containing three pairs of disulfide bonds and adopting a strategy of combining disulfide bond deletion and amino acid site-directed mutagenesis. Furthermore, non-natural amino acid Pen is introduced into the mutant with the optimal activity, so that the stability of the mutant in vivo is remarkably enhanced, and the action time is prolonged.

The invention discloses a human albumin binding peptide 4F1 and a method for purifying human albumin, and belongs to the technical field of polypeptides. The amino acid sequence of the human albumin binding peptide 4F1 comprises an amino acid sequence as shown in SEQ ID NO. 1; and / or an amino acid sequence of a fusion protein with the same function, which is obtained by connecting tag protein to the N terminal and / or C terminal of the amino acid sequence as shown in SEQ ID NO.1. The human albumin binding peptide 4F1 has extremely high affinity with human albumin and can be used for separating and purifying a human albumin solution, and the purity of the purified human albumin far exceeds the pharmacopoeia standard and can reach 99.9% or above. According to the method, high-selectivity capture of affinity chromatography and strong impurity removal capacity of gel filtration chromatography are combined, a purification process route which is efficient, stable and easy to amplify is constructed, and the recovery rate and the production efficiency are remarkably improved.

The present application relates to the technical field of biological medicines, and particularly relates to a polypeptide for specifically blocking the interaction between PD-L1 and endogenous double-stranded RNA and a medicine. The present application provides a polypeptide for specifically binding to endogenous double-stranded RNA, which has a strong effect on the activation of innate immune genes. The polypeptide fused with a cell-penetrating peptide can competitively bind to endogenous double-stranded RNA in cells and inhibit the A-to-I editing of endogenous double-stranded RNA, thereby improving the sensitivity of tumor cells to drugs. The present application provides a new strategy for the immunotherapy of tumor patients and has a wide application prospect.

A nanoagonist, and a preparation method and use thereof are provided, belonging to the technical field of nanoscale biomedicine. The nanoagonist is formed by self-assembly of a transformable peptide, where the transformable peptide includes a targeted antimicrobial peptide, a functionalized self-assembling peptide, an FcγR recognition peptide, and a lipase-responsive hydrophobic molecule that are coupled in sequence. The functionalized self-assembling peptide can control the FcγR recognition peptide to flip toward a surface of a target pathogen during secondary self-assembly, and the target pathogen is a pathogen targeted and bound by the targeted antimicrobial peptide. The nanoagonist combines externalization of the FcγR recognition peptide that can be guided during the secondary self-assembly with FcγR-mediated endocytosis, and a nanoagonist is developed for the first time that takes into account both pathogen clearance and host immune function repair.

The present application provides a Morchella source hypoglycemic peptide and application thereof, and belongs to the technical field of biological medicine. The present application provides a Morchella source hypoglycemic peptide, which comprises a tripeptide and / or a tetrapeptide; the amino acid sequence of the tripeptide is PTW, and the amino acid sequence of the tetrapeptide is MPTW. The Morchella source hypoglycemic peptide of the present application has a molecular weight less than 1000, strong hydrophobicity, and contains Pro and Trp. Molecular docking results show that PTW and / or MPTW can be combined with the proteinase diabetes prevention and treatment target dipeptidyl peptidase IV, protein tyrosine phosphatase 1B and alpha-amylase to play a hypoglycemic role through hydrogen bonding and hydrophobic interaction. Moreover, the hypoglycemic peptide of the present application is a natural product, has no hemolyticity and allergenicity, and has good pharmacokinetic characteristics. The present application has certain guiding significance for the development of food source hypoglycemic peptides.

The application discloses the field of biotechnology, and discloses a kind of nanobody targeting B cell CD20 and application thereof.The amino acid sequence of nanobody complementarity determining region CDR1~CDR3 is selected from SEQ ID NO.1~9.The nanobody provided by the application can specifically bind to B cell in human blood, thereby eliminating B cell producing pathological antibody.The nanobody provided by the application has smaller molecular weight compared with traditional antibody, is stable, low in cost, easy to express and high in expression amount, and is suitable for large-scale production.Nanobody is combined with solid phase carrier in the application, B cell in blood can be targetedly eliminated, the number of B cell in blood of patient is reduced in a short time, antibody level is reduced, thereby improving life quality, reducing complication, and providing new treatment selection for those patients who are ineffective or intolerant to traditional treatment.

The invention discloses a construction method and application of a protein targeted degradation system, and belongs to the technical field of protein targeted degradation. According to the present invention, the Tris 21 Pturbo-GFP recombinant plasmid is constructed, and the Tris 21 Pturbo-GFP recombinant plasmid is combined with the electroporation delivery antibody technology so as to achieve the rapid degradation of the pig endogenous protein, and the dual plasmid co-transfection is combined with the electroporation delivery antibody to achieve the rapid degradation of the foreign protein; compared with traditional siRNA, the protein targeted degradation system has the advantages that the knock-down efficiency is improved by several times or more, and an excellent protein targeted degradation effect is achieved.

The invention provides an anti-MIC humanized single-chain antibody as well as a coding gene, an expression vector, a host, a conjugate or conjugate, an adsorbent and application thereof, and the humanized single-chain antibody provided by the invention can be specifically combined with an NKG2D ligand in human blood or plasma so as to relieve a tumor immunosuppression effect caused by abnormal increase of soluble MIC protein. Compared with the traditional antibody, the humanized single-chain antibody provided by the invention has the advantages of small molecular weight, strong stability, low immunogenicity, low cost, easy expression and high expression quantity, and is suitable for large-scale production. According to the invention, the humanized single-chain antibody is combined with the solid-phase carrier, so that the adsorption efficiency is high, the adsorption performance is stable, the non-specific adsorption capacity on other proteins in plasma is low, the specificity is good, the safety performance in clinical treatment is good, and the economic benefit is high. In addition, the humanized single-chain antibody provided by the invention reduces side effects caused by falling of adsorption ligands, and improves safety.

The invention relates to application of Chinese yam protein, in particular to application of Chinese yam active protein in preparation of medicine for regulating blood testis barrier to treat testis dysfunction. L-YP has unique fluorescence characteristics and remarkable antioxidant activity, and can effectively improve testis immune microenvironment disorder by repairing blood testis barriers and inhibiting NLRP3 and TLR4 / MyD88 / NF-kappa B inflammatory pathways. In combination with a molecular docking technology, a molecular mechanism that key components such as YP2 (mitochondrial ornithine aminotransferase) and the like realize immunoregulation through high-affinity binding is clarified for the first time. According to the application disclosed by the invention, a molecular docking technology is creatively combined with system experimental analysis, and a molecular mechanism of improving testis injury by the Chinese yam protein is illuminated from the perspective of a'structure-function-target 'synergistic effect, so that a novel dietary intervention strategy is provided for preventing reproductive toxicity caused by chemotherapy; the application potential of medicinal and edible components in the field of male reproductive health is highlighted.

The present application relates to the field of biotechnology, in particular to a pair of mutually recognizing and combining alpha helix peptides and application thereof. The alpha helix peptide 1 has an amino acid sequence as shown in SEQ ID No. 1, and the alpha helix peptide 2 has an amino acid sequence as shown in SEQ ID No. 2. The cells are combined with each other by a pair of mutually recognizing and combining alpha helix peptides, so that the cells are arranged in a desired manner. The suitable material concentration and action time are verified; the formed cell group morphology is observed by various microscopes; two kinds of models are successfully constructed; and the improvement of cell function by adding DPH material is verified.

The application discloses a platelet antibody detection and cross matching blocking chromatography kit and a detection method thereof, wherein the kit comprises a U-shaped microwell plate, a chromatography medium for separating platelet immune complexes from non-cell particle components under the action of centrifugal force, freeze-dried platelets, a platelet diluent, a platelet fluorescent staining solution comprising fluoresceinized platelet antibodies specifically labeled for platelets, biotin-labeled anti-human globulin for specifically binding with antibodies bound to the surface of the platelets, avidinized fluorescein for coupling with the biotin-labeled anti-human globulin through a biotin-avidin specific binding system, negative control serum / plasma and positive control serum / plasma. The application combines the blocking chromatography technology with a high-sensitivity flow immunofluorescence method to construct an efficient and accurate platelet antibody detection and cross matching system, which is high in specificity and sensitivity, simple in operation and stable in detection results.

This invention discloses a 2-oxo-1,2-dihydropyridine-3-amide compound, its preparation method, and its uses, belonging to the field of pharmaceutical technology. In the 2-oxo-1,2-dihydropyridine-3-amide compound structure disclosed in this invention, the pyrimidine ring structure can theoretically form hydrogen bond interactions with the receptor, making it more suitable for binding to Mer and c-Met kinases. Furthermore, the novel structural type of compound of this invention exhibits strong enzyme inhibitory activity while also showing specificity and selectivity against tumor cells and normal cells. Simultaneously, the novel structural compound has high safety and fewer toxic side effects, making it easier to use as an anti-tumor drug.

The invention discloses a phage display library panning method for resisting cTnI protein 19C7-39-47 epitopes, and belongs to the technical field of biology. According to the method, eGFP is used as carrier protein, a target epitope sequence for elutriation is fused through linker, an antibody for resisting a target epitope displayed by phage is elutriated through a solid phase, that is, the antibody displayed by the phage combined with the target epitope can be enriched through multiple rounds of elutriation and amplification, only a conventional elisa plate and a constant-temperature shaking table are needed in the process, and the method is simple and convenient. The method has the advantages of low cost, simplicity and convenience in operation and the like, and a new idea is provided for panning of the phage display library.

The application discloses a walnut protein peptide with ADH and ALDH activation activity and a screening method thereof, and belongs to the technical field of food-derived bioactive peptide development. The application creatively combines polypeptidomics, various screening conditions and molecular docking to construct a method for rapidly screening ADH and ALDH activating peptides, thereby avoiding the time-consuming and labor-consuming defects caused by step-by-step separation and purification. The application first identifies a high-activity ADH and ALDH activating polypeptide PGHFEAF from walnut protein, the polypeptide can resist gastrointestinal digestion, the docking and binding energy of the polypeptide with ADH and ALDH are-8.2 and-10.5 kcal / mol respectively, the ADH activation EC 50 value is 0.49+ / -0.09 mM, and the ALDH activation EC 50 value is 1.85+ / -0.28 mM.

The invention discloses a lysosome mediated protein degradation agent based on a circular RNA (Ribonucleic Acid) aptamer and a preparation method of the lysosome mediated protein degradation agent. The annular RNA aptamer is composed of a transferrin receptor RNA aptamer, an RNA aptamer specifically bound with target protein, a fluorescent RNA aptamer bound with a small molecule dye, and RNA ribozyme with two ends capable of spontaneously forming a ring in a human body, wherein the RNA aptamer is composed of a transferrin receptor RNA aptamer, an RNA aptamer specifically bound with target protein, a fluorescent RNA aptamer bound with a small molecule dye, and an RNA ribozyme with two ends capable of spontaneously forming a ring in the human body. The lysosome mediated protein degradation agent based on the circular RNA aptamer has good stability and low biotoxicity, can efficiently degrade target protein, provides a new platform for precise regulation and control and targeted therapy of cells, and has a potential application prospect.

Disclosed in the present invention are antifungal compounds, a preparation method therefor, and the use thereof. The compounds have the following general formula I, wherein L represents a linking moiety, referring to a small molecule fragment, and Ar represents a heterocyclic ring or a substituted heterocyclic ring. The series of compounds of the present invention can better bind to an enzyme target by means of various interaction forces, thereby exhibiting good activity; and the compounds exhibit excellent activity in both in vitro experiments and animal experiments, the activity of most of the compounds against Candida albicans being better than that of fluconazole. In addition, the series of compounds have anti-inflammatory effects while eliminating fungal infections; and compared with voriconazole, the series of compounds of the present invention can remarkably down-regulate the expression of an inflammatory factor interleukin 6.